Differential proteomic analysis of virus-enriched fractions obtained from plasma pools of patients with dengue fever or severe dengue

Abstract

BackgroundDengue is the most widespread mosquito-borne viral disease of public health concern. In some patients, endothelial cell and platelet dysfunction lead to life-threatening hemorrhagic dengue fever or dengue shock syndrome. Prognostication of disease severity is urgently required to improve patient management. The pathogenesis of severe dengue has not been fully elucidated, and the role of host proteins associated with viral particles has received little exploration.MethodsThe proteomes of virion-enriched fractions purified from plasma pools of patients with dengue fever or severe dengue were compared. Virions were purified by ultracentrifugation combined with a water-insoluble polyelectrolyte-based technique. Following in-gel hydrolysis, peptides were analyzed by nano-liquid chromatography coupled to ion trap mass spectrometry and identified using data libraries.ResultsBoth dengue fever and severe dengue viral-enriched fractions contained identifiable viral envelope proteins and host cellular proteins. Canonical pathway analysis revealed the identified host proteins are mainly involved in the coagulation cascade, complement pathway or acute phase response signaling pathway. Some host proteins were over- or under-represented in plasma from patients with severe dengue compared to patients with dengue fever. ELISAs were used to validate differential expression of a selection of identified host proteins in individual plasma samples of patients with dengue fever compared to patients with severe dengue. Among 22 host proteins tested, two could differentiate between dengue fever and severe dengue in two independent cohorts (olfactomedin-4: area under the curve (AUC), 0.958; and platelet factor-4: AUC, 0.836).ConclusionA novel technique of virion-enrichment from plasma has allowed to identify two host proteins that have prognostic value for classifying patients with acute dengue who are more likely to develop a severe dengue. The impact of these host proteins on pathogenicity and disease outcome are discussed.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-015-1271-7) contains supplementary material, which is available to authorized users.

A View of Platelets in Dengue

“…Moreover, dengue patients had increased P-CD62 surface expression on platelets from patients presenting dengue with warning signs and severe dengue syndromes compared to mild dengue. In agreement, they observed exhaustion of the granule-stored chemokine PF4/CXCL4 in platelets from patients with dengue [148], similarly to another study reported that patients with severe dengue have lower levels of PF4/ CXCL4 in plasma when compared to mild dengue patients [149].…”

The Complement System in Flavivirus Infections

“…Another study demonstrated that C1q binds directly to both the DENV E protein and the viral particle, and when C1q was pre-incubated prior to DENV infection of THP-1 cells, decreased viral infectivity was observed ( Douradinha et al, 2014 ). Proteomics data confirmed the binding of several complement proteins, including C1q, C1s, C1r, C5, C9, and mannan-binding lectin serine protease 1 (MASP1) on viral-enriched fractions from DENV-infected patients and on domain III of the DENV E protein ( Fragnoud et al, 2015 ; Huerta et al, 2016 ).…”

Drug repositioning for dengue haemorrhagic fever by integrating multiple omics analyses

“…Fragnoud et al . 43 identified 22 proteins compared proteins purified from plasma pools of dengue fever patients and normal controls. These proteins were analysed by nano-liquid chromatography coupled to ion trap mass spectrometry and identified using data libraries.…”